Detection and molecular characterization of equine herpesviruses 1, 2, and 5 in horses in the Republic of Serbia

Radalj, Andrea; Nišavić, Jakov; Krnjaić, Dejan; Valčić, Miroslav; Jovanović, Tanja; Veljović, Ljubiša; Milić, Nenad

doi:10.2754/avb201887010027

Acta Veterinaria > Archive > 2018, Vol. 87 > Issue 1 > Detection and molecular characterization…

Acta Vet. Brno 2018, 87: 27-34

https://doi.org/10.2754/avb201887010027

Detection and molecular characterization of equine herpesviruses 1, 2, and 5 in horses in the Republic of Serbia

Andrea Radalj¹, Jakov Nišavić¹, Dejan Krnjaić¹, Miroslav Valčić², Tanja Jovanović³, Ljubiša Veljović⁴, Nenad Milić¹

¹University of Belgrade, Faculty of Veterinary Medicine, Department of Microbiology, Belgrade, Serbia
²University of Belgrade, Faculty of Veterinary Medicine, Department of Infectious Diseases of Animals and Diseases of Bees, Belgrade, Serbia
³University of Belgrade, Faculty of Medicine, Institute of Microbiology and Immunology, Virology Department, Belgrade, Serbia
⁴Scientific Veterinary Institute of Serbia, Belgrade, Serbia

Received August 27, 2017
Accepted April 3, 2018

The presence of equine herpesviruses 1, 2 and 5 (EHV-1, EHV-2 and EHV-5) was examined in 66 samples of spinal cord, submandibular lymph nodes and spleen of healthy, non-vaccinated abattoir horses from different locations in the Republic of Serbia. Virus isolation was conducted on RK-13 cell line with the confirmation of isolated viral strains by multiplex nested polymerase chain reaction. The cytopathic effect was observed 48–72 h after the first inoculation in 28 (42.4%) organ samples, and after 5 days in 11 other samples (16.7%) that were all confirmed as EHV-1. Four other samples (6.1%) that showed cytopathic effects on day 5 of the third passage were all positive for EHV-5. Additionally, EHV-1, EHV-2, and EHV-5 were directly detected in all organs by multiplex nested PCR in 46 (69.7%), 3 (4.5%), and 7 (10.6%) samples, respectively. The molecular characterization based on nucleotide sequencing of the part of the gB gene showed that Serbian EHV-1 isolates were 100% homogenous and clustered with EHV-1 strains from Turkey, the United Kingdom, the United States, and Japan. The EHV-2 strain from Serbia branched together with Turkish EHV-2 isolates with homogeneity from 96% to 98%. Serbian EHV-5 strains can be separated in one distinct cluster with isolates from Turkey and the United States with homogeneity from 98 to 99%. These data represent the first report of the molecular characterization of EHV-1, EHV-2, and EHV-5 in the horse population of the Republic of Serbia and document the first successful isolation of Serbian EHV-5 strains.

Keywords

EHV-1, EHV-2, EHV-5, virus isolation, molecular characterization.

Funding

This study was realized within the Project Grant no. TR31008 financed by The Ministry of Education, Science and Technological Development of the Republic of Serbia.